CN109884301B

CN109884301B - Biomarker for survival prognosis prediction of muscle-layer invasive bladder cancer and application of biomarker

Info

Publication number: CN109884301B
Application number: CN201910143394.9A
Authority: CN
Inventors: 蔚青; 毕德玺
Original assignee: Shanghai Tenth Peoples Hospital
Current assignee: Shanghai Tenth Peoples Hospital
Priority date: 2019-02-26
Filing date: 2019-02-26
Publication date: 2023-03-31
Anticipated expiration: 2039-02-26
Also published as: CN109884301A

Abstract

The invention relates to a biomarker for diagnosis and/or prognosis of survival of muscle-layer invasive bladder cancer and application thereof, wherein the biomarker is selected from at least one of GAS6, PDGFC, DDR2, PDGFRA, FN1 and PPARG, and more preferably at least one of two-gene biomarkers GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG and FN1/PPARG. The expression characteristics of the biomarkers in tumor tissues have the value of predicting the survival prognosis of the muscle-layer invasive bladder cancer. The expression of GAS6, PDGFC, DDR2, PDGFRA and FN1 in the tumor and the expression of PPARG are all in negative correlation, and the expression ratio of the PPARG to the expression ratio of the PPARG can be respectively used as independent prognostic indicators. The double-gene molecular characteristic method provided by the invention is simple and easy to implement; the adopted index is the expression ratio of the two markers, and samples can be directly compared without standardization treatment.

Description

Biomarker for survival prognosis prediction of muscle-layer invasive bladder cancer and application of biomarker

Technical Field

The invention belongs to the technical field of biomedicine, and particularly relates to a biomarker for survival prognosis prediction of muscle invasive bladder cancer and application thereof.

Background

Bladder cancer is the most common malignancy of the urinary system. Muscle invasive bladder cancer means that bladder cancer tumor breaks through the basal layer and invades to the muscle layer, the malignancy degree is high, the prognosis is poor, and the 5-year overall survival rate is low. Currently, there is a lack of simple and effective biomarkers for prognosis determination. The use of transcriptomics has greatly facilitated understanding of the molecular characteristics of the disease and its relevance to clinical manifestations. However, the method depends on transcription data, and has complex detection and analysis means, high cost and great difficulty in clinical implementation.

PPARG, peroxisome-activated receptor gamma, belongs to the subfamily of nuclear receptor peroxisome proliferator-activated receptors. PPARG plays an important role in biological processes such as lipid synthesis, carbohydrate metabolism, inflammatory response, and atherosclerosis, and has recently been shown to be associated with various tumors. In bladder cancer, PPARG has been studied to show that it is a potential therapeutic target, and PPARG agonists have a cancer-suppressing effect.

PDGFC, platelet Derived Growth Factor C (Platelet Derived Growth Factor C), belongs to the Platelet Derived Growth Factor family. PDGFC plays an important role in regulation and control in processes such as embryonic development, angiogenesis, cell proliferation and differentiation, cell migration and the like, and recently researches show that PDGFC plays an important role in the growth of tumor cells and the interaction with a tumor microenvironment.

GAS6, growth Arrest Specific gene product 6 (Growth Arrest Specific 6), is a tyrosine kinase receptor Axl, ty-ro3, and Mer ligand that stimulates cell proliferation, cell adhesion, and cell migration. GAS6/AXL signaling plays an important role in a variety of biological processes. GAS6 exhibits high expression in many cancers and is involved in tumor development.

DDR2, namely Discoidin Domain Receptor 2, plays an important role in the interaction process of cells and extracellular microenvironment, is regulated and controlled by extracellular matrix components, is Receptor tyrosine kinase which is closely related to the development process of tumors, and participates in the processes of proliferation, invasion, metastasis and the like of the tumors. Its overexpression is found in urothelial cancer in association with a poor prognosis.

PDGFRA, the platelet-derived growth factor receptor alpha, is a cell surface tyrosine kinase receptor for platelet-derived growth factors. Studies have shown that PDGFRA plays an important role in organ development, wound healing, and tumor progression. Mutational activation is common in gastrointestinal stromal tumors, but the role played in bladder cancer is unknown.

FN1, fibronectin 1 (fibrinectin 1), is a high molecular glycoprotein that is an important component of extracellular matrix, and is involved in biological processes such as cell adhesion and migration, and in cancer metastasis. In bladder cancer, FN1 has been reported to be highly expressed in cancer tissues.

These biomarkers are all likely to be potential bladder cancer tumor markers, but their prognostic role is unknown.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention provides a biomarker for diagnosing and/or predicting survival prognosis of muscle-layer invasive bladder cancer and application thereof, and the expression characteristics of the biomarker in tumor tissues have the value of predicting survival prognosis of muscle-layer invasive bladder cancer.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect of the present invention, there is provided a biomarker for diagnosis and/or prognosis of survival of muscle-invasive bladder cancer, wherein the biomarker is at least one selected from GAS6, PDGFC, DDR2, PDGFRA, FN1, PPARG.

Further, the biomarker is a double-gene biomarker selected from at least one of GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG and FN1/PPARG.

Further, the biomarker is a dual-gene biomarker which is GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG or FN1/PPARG.

The second aspect of the invention provides an application of the biomarker as a diagnosis marker and/or a survival prognosis prediction marker of muscle-layer invasive bladder cancer.

In order to further optimize the application, the technical measures adopted by the invention comprise:

further, the expression ratio of GAS6, PDGFC, DDR2, PDGFRA and FN1 in muscle layer invasive bladder cancer tumor and PPARG can be used as independent prognostic indicators respectively.

Further, the step of calculating the expression ratio of the biomarkers comprises:

step one, processing a specimen to obtain quantitative data of genes; the step is to detect the expression status of the double-gene biomarkers in the tumor tissue specimen, and the detection method is not limited to the detection of the quantitative data, and the detection methods are all conventional detection methods in the field and all follow the standard operation procedures given by the kit.

And step two, calculating the expression ratio of the double-gene biomarkers according to the obtained quantitative data, wherein the double-gene biomarkers are GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG or FN1/PPARG.

Further, the step one is specifically: the RNA-Seq, RT-qPCR or transcriptome chip is adopted by the specimen to obtain the gene transcription quantitative data of the RNA level, or the protein quantitative data is obtained by the specimen through immunohistochemistry.

Further, in the first step, the specimen is a fresh tissue specimen or a formaldehyde-fixed paraffin-embedded specimen.

Further, in step one, the quantitative data for both markers in each of the two-gene biomarkers must be determined using the same method.

Furthermore, the expressions of GAS6, PDGFC, DDR2, PDGFRA and FN1 in the specimen are all in negative correlation with the expression of PPARG, the overall survival is better corresponding to a lower ratio, and the risk of poor prognosis is low.

Compared with the prior art, the invention has the following beneficial effects by adopting the technical scheme:

the invention adopts the molecular characteristics of double genes to judge the prognosis of muscle invasive bladder cancer, only adopts two markers, and has simple method and easy implementation; and the adopted index is the expression ratio of the two markers, and samples can be directly compared without carrying out standardization treatment among the samples. Particularly, the influence caused by different standardization methods can be eliminated aiming at omics data. Compared with the method adopting transcriptomic typing and gene set profile, the method is simpler and more feasible.

Drawings

FIG. 1 is a graph showing the results of correlation analysis of mRNA levels of GAS6, PDGFC, DDR2, PDGFRA, FN1 and PPARG genes based on RNA-Seq data according to an embodiment of the present invention.

FIG. 2 is a graph of the overall survival analysis results of each set of two-gene markers by ratio in one embodiment of the present invention; wherein, L: a low ratio set; m: a set of median comparisons; h: a high ratio set.

FIG. 3 is a graph showing the results of Cox regression analysis of each group of two-gene markers by their inclusion in stage and age factors according to ratios in an embodiment of the present invention; wherein, L: a low ratio group; m: comparing the value group; h: a high ratio set.

FIG. 4 is a graph of the overall survival analysis results of groups of dual gene markers by ratio based on human expression profiling chip data in an embodiment of the present invention; wherein, L: a low ratio set; h: a high ratio set.

Detailed Description

The invention relates to a biomarker for diagnosis and/or prognosis of survival of muscle-layer invasive bladder cancer, which is characterized in that the biomarker is selected from at least one of GAS6, PDGFC, DDR2, PDGFRA, FN1 and PPARG, and more preferably at least one of two-gene biomarkers GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG and FN1/PPARG. The invention also relates to application of the biomarker as a diagnosis marker and/or a survival prognosis prediction marker for the muscle invasive bladder cancer.

The following description of the embodiments of the present invention will be made with reference to the accompanying drawings. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.

Example 1

In this example, 405 RNA-Seq data of muscle-invasive bladder cancer are taken as an example, and the application of two-gene biomarkers GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG, and FN1/PPARG in predicting bladder cancer survival prognosis is as follows:

transcriptome data and patient overall survival data for tumor specimens were downloaded from The Cancer Genome Atlas (TCGA) database (https:// portal. Gdc. Cancer. Gov), with data type FPKM (fragments per foundation of transcript peptides) values.

Expression values of GAS6, PDGFC, DDR2, PDGFRA, FN1 and PPARG were extracted therefrom. The results of Spearman correlation analysis are shown in fig. 1, and it can be seen that the expression values of GAS6, PDGFC, DDR2, PDGFRA, FN1 are all negatively correlated with the expression value of PPARG.

For each group of markers, patients were divided into three groups by gene expression ratio, three digits, a low ratio group, a medium ratio group and a high ratio group, and the grouping threshold of the marker ratios of each group is shown in table 1.

TABLE 1 Fine threshold values for the respective fine marker ratios

	Low specific value group	Middle ratio group	High ratio group
				GAS6/PPARG	＜0.155	Greater than 0.155 and < 1.269	＞1.269
PDGFC/PPARG	＜0.0319	0.0319 and 0.2404	＞0.2404
				DDR2/PPARG	＜0.0199	Greater than 0.0199 and < 0.1587	＞0.1587
PDGFRA/PPARG	＜0.0335	> 0.0335 and < 0.1989	＞0.1989
				FNI/PPARG	＜0.703	> 0.703 and < 8.719	＞8.719

Survival analysis (Kaplan-Meier analysis, log-rank test) was performed on the overall survival of three groups of patients, and the analysis results are shown in FIG. 2, which shows that the overall survival of the low proportion group is significantly better than that of the other two groups.

To confirm that the panel of markers are independent predictors of tumor stage and age of the patient, cox regression analysis was performed with stage and age factors included, using SPSS software operating according to the standard protocol for Cox regression analysis, the results of which are shown in fig. 3, confirming that the ratios of the panel markers can be used as independent prognostic predictors.

In addition, with the FPKM values normalized by quartile, the analysis results were consistent with the above, and it can be seen that the ratio was directly comparable without the need for inter-sample normalization.

Example 2

In this embodiment, using 73 human expression profile chip data of muscle-layer-invasive bladder cancer as an example, the application of the two-gene biomarkers GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG, and FN1/PPARG in predicting the prognosis of bladder cancer survival is as follows:

transcriptome data and patient global survival data for tumor specimens were downloaded from a GEO database (accession number GSE 48277), the data type being human expression profiling chip data.

Similarly to example 1, expression values of PDGFC, DDR2, FN1 and PPARG genes were extracted therefrom, and expression ratios of PDGFC, DDR2, FN1 genes and PPARG genes in each sample were calculated, respectively. And according to the ratio distribution, dividing the ratio into two groups according to median, namely a high ratio group and a low ratio group. Survival analysis of the two groups of patients was performed on the overall survival (Kaplan-Meier analysis, log-rank test) and the results are shown in FIG. 4, showing that the overall survival of patients with low ratios was significantly better than that of patients with high ratios (PDGFC/PPARG: p <0.01, DDR2/PPARG: p <0.01, FN1/PPARG: p = 0.018)

As can be seen from the above examples, the present inventors found that five pairs of biomarkers, namely GAS6/PPARG, PDGFC/PPARG, DDR2/PPARG, PDGFRA/PPARG and FN1/PPARG, are all related to PPARG, and that the expression of GAS6, PDGFC, DDR2, PDGFRA and FN1 in tumors and PPARG are all negatively related, the expression ratio of PPARG and PPARG can be used as independent prognostic indicators, the adopted indicators are the expression ratio of the two markers, the samples can be directly compared without standardization treatment, and the expression characteristics in tumor tissues all have the value of predicting the prognosis of muscle-invasive bladder cancer.

The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, equivalent alterations and modifications are intended to be included within the scope of the present invention, without departing from the spirit and scope of the invention.

Claims

1. Use of a biomarker for the preparation of a product for characterizing the expression level of PPARG in a muscle-invasive bladder cancer patient, wherein the biomarker is selected from at least one of GAS6, PDGFC, DDR2, PDGFRA and FN 1.

2. The use of claim 1, wherein the expression of GAS6, PDGFC, DDR2, PDGFRA, FN1 in the sample is negatively correlated with the expression of PPARG.